乙型肝炎病毒前S1蛋白反式激活蛋白2结合蛋白1基因的克隆及其反式调节基因的筛选

目的 克隆HBV前S1蛋白反式激活蛋白2结合蛋白1(PS1TP2BP1)基因,并筛选与其相互作用的反式激活基因.方法 应用聚合酶链反应(PCR)扩增PS1TP2BP1基因,鉴定正确后再将其亚克隆到真核表达载体pcDNATM3.1/myc-His A上;以真核表达质粒pcDNATM3.1/mycHis A-PS1TP2BP1转染HepG2细胞,构建cDNA消减文库;进行测序及同源性分析.结果 从HepG2细胞来源的cDNA中扩增出PS1TP2BP1基因,并成功进行TA克隆,酶切、测序均正确后,成功构建真核表达重组体.消减文库扩增后得到35个阳性克隆,经菌落PCR分析,得到15个含200～1000 bp插入片段的菌落.对所得片段测序,并进行间源性分析,显示15种已知基因编码蛋白可能是PS1TP2BP1反式激活靶基因.结论 成功克隆PS1TP2BP1,并构建了PS1TP2BP1反式激活基因差异表达的cDNA消减文库,为今后进一步分析、研究病毒蛋白的致病机制奠定了基础.     

机械通气时间对SmartCare/PS脱机功能影响的比较

目的探讨机械通气时间对SmartCare/PS（SC）脱机功能的影响。方法将SC脱机患者分为A（简单脱机）、B（困难脱机、延迟脱机）两组,进行SC实际成功率的比较。对SC提示与实际脱机成功一致组（实际成功组）和SC提示成功但实际脱机失败组（实际失败组）的呼吸力学指标进行比较。结果两组SC实际脱机成功率有统计学意义（P〈0.01）。SC实际失败组的RR快,Vt、MIP等均显著低于SC实际成功组;RSBI各时间段等均高于SC实际成功组,差异有统计学意义（P〈0.01）。结论 SC应用于简单脱机准确率高,而对困难脱机、延迟脱机应用SC脱机准确性偏低,需结合MIP、RSBI的动态变化以提高实际脱机的成功率。     

肺表面活性物质治疗新生儿肺透明膜病的临床分析

目的探讨肺表面活性物质治疗新生儿肺透明膜病的临床价值。方法随机性的抽取了我院2009年6月至2010年6月收治的48例肺透明膜病患儿,分为研究组和对照组各24例。研究组在出生后1d内使用肺表面活性物质来治疗,对照组不使用。比较X线、并发症和呼吸时间,评测肺表面活性物质疗效。结果研究组和对照组在X线、并发症和呼吸时间等有明显差异,卡方检验P<0.05。结论肺表面活性物质在治疗新生儿肺透明膜病上有较好的疗效,能减少并发症的发生,还能减少新生儿病死率。     

Interdependence of measures in pavlovian conditioned freezing

Pavlovian conditioned freezing is an intensively utilized paradigm that has become a standard model of memory and cognition. Despite its widespread use, the interdependence among each measure commonly reported in fear conditioning studies has not been described. Using mice, we examine the relationship of each common freezing measure (Training Baseline, Post-Shock freezing, Contextual Fear, Tone Baseline, and Tone Fear), as well as baseline locomotor activity measures, to better understand the significance of each. Of particular interest, Post-Shock freezing appears to be a good measure of immediate contextual memory. In contrast, Tone Baseline freezing, as typically measured in a novel context, appears to be contaminated with multiple sources of fear. Finally, Contextual and Tone Fear show a weak interdependence.     

Evolution of neuronal patterning in free-living rhabditid nematodes I: Sex-specific serotonin-containing neurons

As a first step toward understanding the evolution of neuronal patterning and function in a group of simple animals, we have examined serotonin-containing neurons in 17 species of free-living rhabditid nematodes and compared them with identified neurons of Caenorhabditis elegans. We found many serotonin-immunoreactive (serotonin-IR) neurons that are likely homologs of those in C. elegans; this paper focuses on sex-specific neurons such as the egg laying hermaphrodite-specific neurons (HSNs), VCs, and male CAs, CPs, and ray sensory neurons known to function in mating. These cells vary in number and position in the species examined but are consistent with a current molecularly based phylogeny. Two groups (Oscheius and Pristionchus) appear independently to have lost a serotonin-IR HSN. Oscheius furthermore has no serotonin-IR innervation of the vulval region, in contrast to every other species we examined. We also saw variation in the location of somas of putative HSN, consistent with evolutionary changes in HSN migration. In C. elegans, the HSN soma migrates during embryogenesis from the tail to the central body, where it innervates its major postsynaptic targets, the vulval muscles. For other species, we observed putative HSN homologs along the anterior-posterior axis from the head to the tail, but typically HSNs were located near the vulva, which also varies in anterior-posterior position among the species we examined. The varying positions of the HSN somas in other species are reminiscent of phenotypes seen in various C. elegans mutants with altered HSN migration, suggesting possible mechanisms for the evolutionary differences we observed.     

The treatment of venous thromboembolism in special populations

Anticoagulant therapy for the typical venous thromboembolism patient is straightforward with predictably favorable outcomes. However, for certain patients with venous thromboembolism, there remains uncertainty and controversy about optimal treatment. These controversial areas include venous thromboembolism patients with: heparin resistance, renal insufficiency, morbid obesity, cancer, antiphospholipid antibody syndrome, recurrent thrombosis despite appropriate anticoagulation, and patients with unprovoked VTE who may or may not benefit from thrombophilia testing. This review summarizes the current data for these special patient populations with venous thromboembolism and provides our recommendations for management.     

Haplotype of thrombomodulin gene associated with plasma thrombomodulin level and deep vein thrombosis in the Japanese population

INTRODUCTION: Thrombomodulin (TM) is an essential cofactor in protein C activation by thrombin. Here, we evaluated the contribution of genetic variations in the TM gene to soluble TM (sTM) level and deep vein thrombosis (DVT) in Japanese. PATIENTS AND METHODS: We sequenced the TM putative promoter, exon, and 3'-untranslated region in DVT patients (n=118). Among 17 genetic variations we identified, two missense mutations (R385K, D468Y) and three common single nucleotide polymorphisms (-202G>A, 2487A>T, 2729A>C) were genotyped in a general population of 2247 subjects (1032 men and 1215 women) whose sTM levels were measured. We then compared the frequency of these mutations in DVT patients with that in the age, body mass index-adjusted population-based controls. RESULTS: We identified one neutral mutation (H381) and three missense mutations (R385K; n=2, A455V; n=53 heterozygous, n=14 homozygous, D468Y; n=2) of TM in the DVT patients. Age-adjusted mean values of sTM were lower in C-allele carriers of 2729A>C than in noncarriers in the Japanese general population (women: 16.7+/-0.3 U/ml vs. 17.9+/-0.2 U/ml, p<0.01, men: 19.4+/-0.3 U/ml vs. 20.4+/-0.3 U/ml, p=0.03). Additionally, the CC genotype of this mutation was more common in the male DVT patients than in the male individuals of the general population (odds ratio=2.76, 95% confidence interval=1.14-6.67; p=0.02). This mutation was in linkage disequilibrium (r-square>0.9) with A455V mutation. CONCLUSIONS: TM mutations, especially those with a haplotype consisting of 2729A>C and A455V missense mutation, affect sTM levels, and may be associated with DVT in Japanese.